A novel isothermal whole genome sequencing approach for Monkeypox Virus
Abstract Monkeypox virus (MPXV) is the zoonotic agent responsible for mpox, an often-self-limiting pox-like disease. Since May 2022, an outbreak characterized by increased human-to-human transmission was detected outside the endemic regions. Whole genome sequencing (WGS) has been successfully used t...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-09-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-024-73613-3 |
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| author | Matthias Licheri Manon Flore Licheri Lukas Probst Cora Sägesser Pascal Bittel Franziska Suter-Riniker Ronald Dijkman |
| author_facet | Matthias Licheri Manon Flore Licheri Lukas Probst Cora Sägesser Pascal Bittel Franziska Suter-Riniker Ronald Dijkman |
| author_sort | Matthias Licheri |
| collection | DOAJ |
| description | Abstract Monkeypox virus (MPXV) is the zoonotic agent responsible for mpox, an often-self-limiting pox-like disease. Since May 2022, an outbreak characterized by increased human-to-human transmission was detected outside the endemic regions. Whole genome sequencing (WGS) has been successfully used to keep track of viral evolution during outbreaks or for surveillance of multiple pathogens of public health significance. Current WGS protocols for MPXV are either based on metagenomic sequencing or tiled-PCR amplification. The latter allows multiplexing due to the efficient enrichment of the viral DNA, however, mutations or the presence of different clades can negatively influence genome coverage yield. Here, we present the establishment of a novel isothermal WGS method for MPXV based on Phi29 DNA polymerase-based multiple displacement amplification (MDA) properties making use of only 6 primers. This approach yielded from 88% up to 100% genome coverage using either alkaline denatured extracted DNA or clinical material as starting material, with the highest coverage generated by clinical material. We demonstrate that this novel isothermal WGS protocol is suitable for monitoring viral evolution during MPXV outbreaks and surveillance in any conventional laboratory setting. |
| format | Article |
| id | doaj-art-d9c9d72e456e4cd1be378e91b48fedc9 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-d9c9d72e456e4cd1be378e91b48fedc92025-02-09T12:37:59ZengNature PortfolioScientific Reports2045-23222024-09-0114111110.1038/s41598-024-73613-3A novel isothermal whole genome sequencing approach for Monkeypox VirusMatthias Licheri0Manon Flore Licheri1Lukas Probst2Cora Sägesser3Pascal Bittel4Franziska Suter-Riniker5Ronald Dijkman6Institute for Infectious Diseases, University of BernInstitute for Infectious Diseases, University of BernInstitute for Infectious Diseases, University of BernInstitute for Infectious Diseases, University of BernInstitute for Infectious Diseases, University of BernInstitute for Infectious Diseases, University of BernInstitute for Infectious Diseases, University of BernAbstract Monkeypox virus (MPXV) is the zoonotic agent responsible for mpox, an often-self-limiting pox-like disease. Since May 2022, an outbreak characterized by increased human-to-human transmission was detected outside the endemic regions. Whole genome sequencing (WGS) has been successfully used to keep track of viral evolution during outbreaks or for surveillance of multiple pathogens of public health significance. Current WGS protocols for MPXV are either based on metagenomic sequencing or tiled-PCR amplification. The latter allows multiplexing due to the efficient enrichment of the viral DNA, however, mutations or the presence of different clades can negatively influence genome coverage yield. Here, we present the establishment of a novel isothermal WGS method for MPXV based on Phi29 DNA polymerase-based multiple displacement amplification (MDA) properties making use of only 6 primers. This approach yielded from 88% up to 100% genome coverage using either alkaline denatured extracted DNA or clinical material as starting material, with the highest coverage generated by clinical material. We demonstrate that this novel isothermal WGS protocol is suitable for monitoring viral evolution during MPXV outbreaks and surveillance in any conventional laboratory setting.https://doi.org/10.1038/s41598-024-73613-3Monkeypox virusMpoxWhole-genome sequencingMDA |
| spellingShingle | Matthias Licheri Manon Flore Licheri Lukas Probst Cora Sägesser Pascal Bittel Franziska Suter-Riniker Ronald Dijkman A novel isothermal whole genome sequencing approach for Monkeypox Virus Scientific Reports Monkeypox virus Mpox Whole-genome sequencing MDA |
| title | A novel isothermal whole genome sequencing approach for Monkeypox Virus |
| title_full | A novel isothermal whole genome sequencing approach for Monkeypox Virus |
| title_fullStr | A novel isothermal whole genome sequencing approach for Monkeypox Virus |
| title_full_unstemmed | A novel isothermal whole genome sequencing approach for Monkeypox Virus |
| title_short | A novel isothermal whole genome sequencing approach for Monkeypox Virus |
| title_sort | novel isothermal whole genome sequencing approach for monkeypox virus |
| topic | Monkeypox virus Mpox Whole-genome sequencing MDA |
| url | https://doi.org/10.1038/s41598-024-73613-3 |
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